title: Chronic cadmium exposure in vitro induces cancer cell characteristics in human lung cells.
abstract: Cadmium is a known human lung carcinogen. Here, we attempt to develop an in vitro model of cadmium-induced human lung carcinogenesis by chronically exposing the peripheral lung epithelia cell line, HPL-1D, to a low level of cadmium. Cells were chronically exposed to 5 μM cadmium, a noncytotoxic level, and monitored for acquired cancer characteristics. By 20 weeks of continuous cadmium exposure, these chronic cadmium treated lung (CCT-LC) cells showed marked increases in secreted MMP-2 activity (3.5-fold), invasion (3.4-fold), and colony formation in soft agar (2-fold). CCT-LC cells were hyperproliferative, grew well in serum-free media, and overexpressed cyclin D1. The CCT-LC cells also showed decreased expression of the tumor suppressor genes p16 and SLC38A3 at the protein levels. Also consistent with an acquired cancer cell phenotype, CCT-LC cells showed increased expression of the oncoproteins K-RAS and N-RAS as well as the epithelial-to-mesenchymal transition marker protein Vimentin. Metallothionein (MT) expression is increased by cadmium, and is typically overexpressed in human lung cancers. The major MT isoforms, MT-1A and MT-2A were elevated in CCT-LC cells. Oxidant adaptive response genes HO-1 and HIF-1A were also activated in CCT-LC cells. Expression of the metal transport genes ZNT-1, ZNT-5, and ZIP-8 increased in CCT-LC cells culminating in reduced cadmium accumulation, suggesting adaptation to the metal. Overall, these data suggest that exposure of human lung epithelial cells to cadmium causes acquisition of cancer cell characteristics. Furthermore, transformation occurs despite the cell's ability to adapt to chronic cadmium exposure.
authors.affiliation: Inorganic Toxicology Group, National Toxicology Program Laboratory, Division of the National Toxicology Program, National Institute of Environmental Health Sciences, Research Triangle Park, NC, USA.
authors.forename: Rachel J
authors.initials: RJ
authors.lastname: Person
authors.forename: Erik J
authors.initials: EJ
authors.lastname: Tokar
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Xu
authors.forename: Ruben
authors.initials: R
authors.lastname: Orihuela
authors.forename: Ntube N Olive
authors.initials: NN
authors.lastname: Ngalame
authors.forename: Michael P
authors.initials: MP
authors.lastname: Waalkes
mesh_terms: D002104:Cadmium / Q000008:administration & dosage / Q000633:toxicity*; D002471:Cell Transformation, Neoplastic / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D004249:DNA Damage / Q000187:drug effects / Q000502:physiology; D006801:Humans; D008168:Lung / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008175:Lung Neoplasms / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D013997:Time Factors; D014407:Tumor Cells, Cultured
chemical_list: D002104:Cadmium
keywords: 4′,6-diamidino-2-phenylindole; 5,5-dimethyl-1-pyrroline N-oxide; Adaptation; BSA; CCT-LC; Cadmium; DAPI; DMPO; Epithelial-to-mesenchymal transition; HIF-1A; HO-1; HPL; Human lung cells; INT; IST; LC50; LUCA; Lung cancer; MMP-2; MT-1A; MT-2A; PBS; Transformation; ZIP; ZNT; Zinc Transporter; Zrt/Irt-like Proteins; bovine serum albumin; chronic cadmium treated-lung cells; heme oxygenase-1; human peripheral lung; hypoxia inducible factor-1 alpha; immuno-spin trapping; lethal concentration 50; lung cancer; matrix metalloproteinase-2; metallothionein-1A; metallothionein-2A; p-iodonitro-tetrazolium violet; phosphate buffered saline; qRT-PCR; quantitative real time reverse transcription polymerase chain reaction